Volumetric analysis of epithelial morphogenesis with high spatiotemporal resolution

高时空分辨率上皮形态发生的体积分析

• 批准号: 10586534
• 负责人: James Todd Blankenship
• 金额: $ 29.49万
• 依托单位: UNIVERSITY OF DENVER (COLORADO SEMINARY)
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10586534
• 关键词: 3-Dimensional; Actins; Actomyosin; Address; Adhesives; Adopted; Adult; Affect; Apical; Architecture; Area; Automobile Driving; Behavior; Biological Models; Cell Aging; Cell Shape; Cell Volumes; Cell surface; Cells; Cellular biology; Columnar Epithelium; Computer Analysis; Coupled; Cytoplasm; Data; Data Set; Development; Dimensions; Disease; Drosophila genus; Elasticity; Embryo; Environment; Epithelial Cells; Epithelium; Event; F-Actin; Gastrula; Generations; Goals; Growth; Homeostasis; Image; Individual; Intercalated Cell; Knowledge; Lateral; Life; Light; Location; Maps; Measurement; Measures; Mechanics; Methods; Microscopy; Molecular; Molecular Conformation; Morphogenesis; Morphology; Movement; Myosin ATPase; Myosin Type II; Nature; Population; Population Distributions; Positioning Attribute; Process; Proteins; Resolution; Shapes; Speed; Stratum Basale; Surface; Surveys; System; Testing; Three-dimensional analysis; Time; Tissues; Viscosity; cell behavior; cell cortex; cell dimension; cell motility; cellular imaging; computerized tools; embryo cell; fly; gastrulation; imaging capabilities; in vivo; intercalation; novel; spatiotemporal; tool

Project Summary How epithelial sheets remodel themselves to adopt new tissue conformations through changes in neighbor relationships and cell shape dynamics has been a key question in development and disease. Interestingly, many of the pioneering studies performed in model systems have largely been confined to 2D analysis, and have often been challenged to image cell behaviors that occur in basal regions that lie deeper into the tissue. The intercalation movements that occur during tissue elongation in the Drosophila gastrula have been a classic system for understanding epithelial remodeling, and have been fundamental to informing the developmental paradigms that describe how cells can change position in an adherent epithelium. Nearly all of the studies in this system have been confined to 2D analysis of apical events in the early fly embryo, and no studies to date have systematically analyzed the full 3D behaviors that drive epithelial remodeling and tissue extension in the Drosophila embryonic epithelium. Thus, one of the biggest remaining questions in the field is how the volumetric nature of epithelial cells affects force propagation and remodeling of the cell surface along the entire apical-basal axis. Fundamental questions on where forces originate from as well as how far and fast forces propagate across different apical-basal layers have remained unanswered. In our preliminary analysis, we have been successful in completing the first full 4D segmentation of the intercalating Drosophila epithelium through the use of Lattice Light Sheet Microscopy (LLSM). We find that intercalation can be initiated at any position we have surveyed along the apical-basal axis. This is striking as previous studies have largely implicated apical force generation, and a single study has suggested that contractile forces can also originate from the basal surface of the epithelium. In the proposed project, we are developing the tools to perform the first comprehensive, quantitative 3D analysis of cell intercalation in the early Drosophila embryo. We will then determine the molecular mechanisms driving 3D force generation, and whether different mechanical regimes exist across the apical-basal axis. Preliminary data suggests highly novel dynamic Myosin II and F-actin populations that show rapid axial propagation in lateral and basal regions. The 3D distributions of these populations are being mapped and the relevant actin nucleating and Myosin regulatory networks will be determined. These results will provide the first comprehensive understanding of the cortical and contractile networks that determine the mechanical environment of a gastrulating epithelium. We will also use 3D data sets in wild-type and functionally compromised backgrounds to examine how epithelial forces propagate along apical-basal and planar dimensions using topological mapping metrics. We will determine how far, and at what velocities, contractile forces spread in an intact, developing epithelium. These results will give fundamental answers into how the viscous cytoplasm and elastic cell cortex respond to force-driven displacements, and how these displacements spread within individual cells and across tissues to drive new tissue topologies.

项目概要 上皮片如何通过邻近的变化来重塑自身以采用新的组织构象 关系和细胞形状动力学一直是发育和疾病的关键问题。有趣的是， 在模型系统中进行的许多开创性研究很大程度上局限于二维分析，并且 经常面临对组织深处基底区域中发生的细胞行为进行成像的挑战。 果蝇原肠胚组织伸长过程中发生的嵌入运动是经典的 系统用于理解上皮重塑，并且对于通知发育至关重要 描述细胞如何改变粘附上皮中位置的范例。几乎所有的研究都在 该系统仅限于早期果蝇胚胎顶端事件的二维分析，迄今为止还没有研究 系统地分析了驱动上皮重塑和组织延伸的完整 3D 行为 果蝇胚胎上皮。因此，该领域剩下的最大问题之一是如何 上皮细胞的体积性质影响整个细胞表面的力传播和重塑 顶底轴。关于力的来源以及力的距离和速度的基本问题 在不同的顶基底层传播仍然没有答案。在我们的初步分析中，我们 已成功完成第一个完整的嵌入果蝇上皮的 4D 分割 通过使用晶格光片显微镜 (LLSM)。我们发现插层可以在任何时刻开始 我们沿着顶底轴测量的位置。这是惊人的，因为之前的研究很大程度上 涉及心尖力的产生，一项研究表明收缩力也可以产生 来自上皮的基底表面。在拟议的项目中，我们正在开发工具来执行 首次对早期果蝇胚胎中的细胞嵌入进行全面、定量的 3D 分析。我们随后将 确定驱动 3D 力生成的分子机制，以及不同的机械机制是否 存在于顶底轴上。初步数据表明高度新颖的动态肌球蛋白 II 和 F-肌动蛋白 在侧面和基部区域表现出快速轴向传播的种群。这些的 3D 分布 正在绘制种群图，并将绘制相关的肌动蛋白成核和肌球蛋白调节网络 决定。这些结果将提供对皮质和收缩的首次全面了解 决定原肠胚上皮机械环境的网络。我们还将使用 3D 数据 设置在野生型和功能受损的背景中，以检查上皮力如何传播 使用拓扑映射度量的顶端-基底和平面尺寸。我们将确定距离和时间 速度、收缩力在完整、发育的上皮中传播。这些结果将为基础 回答粘性细胞质和弹性细胞皮层如何响应力驱动的位移，以及 这些位移如何在单个细胞内和跨组织传播以驱动新的组织拓扑。